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Amount of σA bound to E after competition with σB and vice versa. Mixtures of 0.1 μM His6-σA and 0.1 μM E (A) or 4 μM His6-σB and 0.1 μM E (B) were incubated before the addition of increasing amounts of His6-σB or His6-σA, respectively. Proteins were analyzed by native PAGE, Western blotting, and immunodetection with His6-σA-specific polyclonal antibodies. The amount of σA in the holoenzyme is shown.
